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. 2011 Jul 28;6(7):e22773. doi: 10.1371/journal.pone.0022773

Figure 1. Distinguishing adherent and internalized cryptococci.

Figure 1

Panels A–C, J774.16 cells were exposed to fungi labeled with anti-capsular mAb and adherent fungi were labeled with an Alexa Fluor 546-labeled secondary antibody (red). Host cells were then permeabilized, and the samples were restained with an Alexa Fluor 488-labeled secondary antibody to stain all fungi (green). Panel A, red channel; Panel B, green channel; Panel C, merge with DIC and DAPI-stained images. Panels D–E, Confocal images of THP-1 cells were challenged for 1 hr with Lucifer Yellow-stained fungi and stained with CellMask (pink). All fungi appear green in the merged image (D), but only engulfed fungi exclude the cytosolic dye and appear as dark silhouettes in the pink channel (E). Scale bars, 5 µm.